title: E1- and ubiquitin-like proteins provide a direct link between protein conjugation and sulfur transfer in archaea.
abstract: Based on our recent work with Haloferax volcanii, ubiquitin-like (Ubl) proteins (SAMP1 and SAMP2) are known to be covalently attached to proteins in archaea. Here, we investigated the enzymes required for the formation of these Ubl-protein conjugates (SAMPylation) and whether this system is linked to sulfur transfer. Markerless in-frame deletions were generated in H. volcanii target genes. The mutants were examined for: (i) the formation of Ubl protein conjugates, (ii) growth under various conditions, including those requiring the synthesis of the sulfur-containing molybdenum cofactor (MoCo), and (iii) the thiolation of tRNA. With this approach we found that UbaA of the E1/MoeB/ThiF superfamily was required for the formation of both SAMP1- and SAMP2-protein conjugates. In addition, UbaA, SAMP1, and MoaE (a homolog of the large subunit of molybdopterin synthase) were essential for MoCo-dependent dimethyl sulfoxide reductase activity, suggesting that these proteins function in MoCo-biosynthesis. UbaA and SAMP2 were also crucial for optimal growth at high temperature and the thiolation of tRNA. Based on these results, we propose a working model for archaea in which the E1-like UbaA can activate multiple Ubl SAMPs for protein conjugation as well as for sulfur transfer. In sulfur transfer, SAMP1 and SAMP2 appear specific for MoCo biosynthesis and the thiolation of tRNA, respectively. Overall, this study provides a fundamental insight into the diverse cellular functions of the Ubl system.
authors.affiliation: Department of Microbiology and Cell Science, University of Florida, Gainesville, FL 32611-0700, USA.
authors.forename: Hugo V
authors.initials: HV
authors.lastname: Miranda
authors.forename: Nikita
authors.initials: N
authors.lastname: Nembhard
authors.forename: Dan
authors.initials: D
authors.lastname: Su
authors.forename: Nathaniel
authors.initials: N
authors.lastname: Hepowit
authors.forename: David J
authors.initials: DJ
authors.lastname: Krause
authors.forename: Jonathan R
authors.initials: JR
authors.lastname: Pritz
authors.forename: Cortlin
authors.initials: C
authors.lastname: Phillips
authors.forename: Dieter
authors.initials: D
authors.lastname: Söll
authors.forename: Julie A
authors.initials: JA
authors.lastname: Maupin-Furlow
mesh_terms: D000595:Amino Acid Sequence; D019843:Archaeal Proteins / Q000737:chemistry / Q000378:metabolism*; D055786:Gene Knockout Techniques; D019766:Genes, Archaeal / Q000235:genetics; D019622:Haloferax volcanii / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D008954:Models, Biological; D008969:Molecular Sequence Data; D008982:Molybdenum / Q000378:metabolism; D012343:RNA, Transfer / Q000378:metabolism; D013438:Sulfhydryl Compounds / Q000378:metabolism; D013455:Sulfur / Q000378:metabolism*; D013696:Temperature; D044764:Ubiquitin-Activating Enzymes / Q000378:metabolism*; D054875:Ubiquitination; D014452:Ubiquitins / Q000378:metabolism*
chemical_list: D019843:Archaeal Proteins; D013438:Sulfhydryl Compounds; D014452:Ubiquitins; D013455:Sulfur; D008982:Molybdenum; D012343:RNA, Transfer; D044764:Ubiquitin-Activating Enzymes